11 results about "Mgb probe" patented technology

The invention discloses a composition, a kit and a method for detecting a mitochondrial gene of sea eel. A primer probe composition targeting the D-LOOP gene of the sea eel mitochondria is designed, and the nucleotide sequences of the primer probe composition correspond to SEQ ID No.1-3. On the basis, the invention further constructs and optimizes a specific real-time fluorescent PCR detection method for the D-LOOP gene of the sea eel mitochondria. According to the method, a part of sequence of the D-LOOP gene of the mitochondria of the sea eel is taken as a target region, a primer and a TaqMan-MGB probe are designed in a targeted manner, and finally a set of rapid, sensitive and high-specificity detection system is established. The detection system can be effectively applied to adulteration and germplasm identification work of the sea eel, and has a very wide industrial application prospect.

The application discloses primers and a probe for real-time fluorescent quantitative PCR detection of a porcine epidemic diarrhea virus G2c subtype and application thereof and belongs to the field of animal virus molecular biology testing methods. The application compares S genes of different gene subtype strains of the porcine epidemic diarrhea virus, designs a pair of primers and a Taqman-MGB probe on the basis of a mutation site specific to the G2c subtype strain, and proves that the primers and the probe can effectively distinguish the G2c and other PEDV gene subtype strains through virus liquids and clinical samples of different gene subtype strains of the PEDV, meanwhile, the primers and the probe have no cross reaction to other viruses of the porcine enteric diarrhea syndrome, and the detection sensitivity of the kit can reach 5 copies / muL. The application can realize specific and sensitive detection of the G2c subtype, and has a positive significance for diagnosis and epidemiological investigation of the porcine epidemic diarrhea virus.

The invention discloses a primer probe set for identifying coreius guichenoti and application. The primer probe group comprises an upstream primer COX1-F1 (SEQ ID NO. 1), a downstream primer COX1-R1 (SEQ ID NO. 2), and a TaqMan-MGB probe COX1-P (SEQ ID NO. 3), and is characterized in that the upstream primer COX1-F1 (SEQ ID NO. 1), the downstream primer COX1-R1 (SEQ ID NO. 2) and the TaqMan-MGB probe COX1-P (SEQ ID NO. Experiments prove that the primer probe group has extremely high specificity on coreius guichenoti, and has no cross reaction with other common fishes which belong to the same genus and are distributed in the same watershed; the qPCR detection method is high in sensitivity, the lowest detection limit can reach 10 copies / mu L, and the linearity of a standard curve is excellent (Rgt; the invention provides a high-sensitivity and high-specificity molecular detection tool and a related detection method for rapid species identification, resource monitoring and endangered mechanism research of coreius guichenoti, and particularly has a wide application prospect in the aspect of non-invasive environmental DNA monitoring.

The invention discloses a detection method for noninvasive prenatal detection of fetal trisomy 21 syndrome based on digital PCR, and relates to the technical field of digital PCR. According to the invention, a target area detection amplification primer and a Taqman-MGB probe are designed according to different sequences of human chromosome 21 genes, and a specific common amplification primer and a specific Taqman-MGB probe are designed according to similar sequences of the human chromosome 21 genes. Accurate detection of a low-proportion trisomy 21 syndrome positive sample is realized, and the omission ratio is greatly reduced. Compared with next-generation sequencing, multiple qPCR technologies and the like, the detection method provided by the invention has the advantages of simplicity in operation, simplicity in data analysis, short detection period, low cost and the like, meanwhile, by innovatively using a similar sequence design technology, high consistency of amplification efficiency of a detection gene and a reference gene is ensured, and the detection accuracy is improved. The detection sensitivity and accuracy are obviously improved.

The invention relates to a primer and a TaqMan-MGB probe group for fluorescent quantitative PCR (Polymerase Chain Reaction) detection of virulent and attenuated strains of classical waterfowl parvovirus, the primer and the TaqMan-MGB probe group comprise a fluorescent quantitative PCR detection primer and a probe group for virulent and attenuated strains of goose parvovirus respectively, and the sequences of the primer and the probe group are shown as SEQ ID NO.1-5 respectively; the primer and the TaqMan-MGB probe group further comprise a fluorescent quantitative PCR detection primer and a probe group for virulent and attenuated muscovy duck parvovirus respectively, and the sequences of the primer and the probe group are shown as SEQ ID NO.6-9 respectively. According to the invention, through systematic test verification and reaction condition optimization, an optimal primer-probe combination is screened out, a GPV TaqMan-MGB dual fluorescent quantitative PCR detection method and an MDPV TaqMan dual fluorescent quantitative PCR detection method are respectively and successfully established, and the detection of virulent and attenuated classical waterfowl parvovirus is realized.

The application belongs to the technical field of microorganism detection, and particularly relates to detection primers and probes for Euryphallus sinensis and application thereof. The application establishes a real-time fluorescent quantitative PCR method for the pathogen based on TaqMan-MGB probes according to the small subunit ribosomal RNA (SSUrRNA) gene of Euryphallus sinensis, and designs detection primers based on isothermal amplification technology. With the constructed recombinant plasmid standard as a template, the detection sensitivity of the quantitative PCR detection method for the target gene is 9.58 copies / reaction. The detection primers for isothermal amplification can be detected under simple constant temperature conditions, and the color change of the detection product after adding nucleic acid dye can realize on-site rapid diagnosis of the pathogen.

The invention discloses a screening method and application of blast-resistant rice, relates to the technical field of nucleotide molecule screening, and in particular relates to a screening method of blast-resistant rice and a TaqMan-MGB probe for specifically detecting a blast-resistant gene Pigm of rice. The method can be used for performing gene molecule on a plurality of rice samples in a high-throughput and high-efficiency manner and automatically screening out the rice with the homozygous Pigm genotype, so that the effect of quickly screening the rice blast-resistant rice is realized.

The application relates to the technical field of molecular biology detection, and discloses a z. laticeps environmental DNA detection kit, which comprises a primer probe composition for real-time fluorescent PCR detection designed based on a z. laticeps mitochondrial Cytb gene, the composition comprises an upstream primer, a downstream primer and a TaqMan-MGB probe, the nucleotide sequence of the upstream primer is shown in SEQ ID No. 1, the nucleotide sequence of the downstream primer is shown in SEQ ID No. 2, and the nucleotide sequence of the TaqMan-MGB probe is shown in SEQ ID No. 3; the 5' end of the TaqMan-MGB probe is modified with a fluorescent reporter group, and the 3' end is coupled with a small groove binder and a non-fluorescent quenching group; the application creates a special primer probe composition and a complete detection system for z. laticeps environmental DNA detection, realizes high specificity and high sensitivity detection of z. laticeps, and uses z. laticeps specific mitochondrial genes as detection targets; the TaqMan-MGB probe can accurately identify slight differences in gene sequences and effectively distinguish z. laticeps from closely related species in the same region.

The invention discloses a composition, a kit and a method for detecting Huso dauricus mitochondrial genes. A specific primer probe composition is designed aiming at the Huso dauricus mitochondrial ND4 gene, and the corresponding nucleotide sequences of the specific primer probe composition are SEQ ID No.1-3. By taking the Huso dauricus mitochondrial ND4 gene as a core technical support, the invention further constructs and optimizes a real-time fluorescent PCR specific detection scheme based on the Huso dauricus mitochondrial ND4 gene. According to the scheme, a part of sequence of a Huso dauricus mitochondrial ND4 gene is selected as a target fragment, a primer and a TaqMan-MGB probe are directionally designed, and finally a set of detection system with rapidness, high sensitivity and strong specificity is successfully established. The detection system can be effectively applied to adulteration identification and germplasm resource identification of huso dauricus products, and has very wide industrial popularization and application prospects.

The present application relates to the technical field of nucleotide molecule screening, in particular to a mutagenesis screening method and application of a rice herbicide-resistant genotype. The present application designs a qPCR reaction system based on a specific SNP site region of a mutant ALS gene, realizes the effect of high-throughput and large-scale screening of a rice parent containing a mutant ALS gene, and can significantly improve the efficiency of rice screening and breeding. The present application designs a TaqMan-MGB probe based on a specific SNP site region of a mutant ALS gene, which can form a hybrid molecule with a specific SNP site with high specificity and high accuracy, ensuring the accuracy and reproducibility of qPCR.

The invention discloses primers and a probe for real-time fluorescent quantitative PCR (polymerase chain reaction) detection of porcine epidemic diarrhea virus G2c subtype and application of the primers and the probe, and belongs to the field of animal virus molecular biology detection methods. By comparing S genes of different gene subtype strains of the porcine epidemic diarrhea virus, a pair of primers and a Taqman-MGB probe are designed on the basis of specific mutation sites of the G2c subtype strain, through virus solutions and clinical samples of the different gene subtype strains of the PEDV, it is proved that the primers can effectively distinguish G2c from other PEDV gene subtype strains, and the G2c and other PEDV gene subtype strains can be effectively distinguished. Meanwhile, no cross reaction exists on other viruses of the porcine intestinal diarrhea syndrome, and the detection sensitivity of the kit can reach 5 copies / microliter. According to the invention, the specific and sensitive detection of the G2c subtype can be realized, and the kit has positive significance on the diagnosis and epidemiological investigation of the porcine epidemic diarrhea virus.